Variation in H&E staining is often a confounder when comparing data in multi-site studies, and impacts on the generalisability of AI models. In this paper, slides containing serial sections of control liver tissue were sent out to eight clinical laboratories in England to stain over a two-week period. H&E staining optical density was quantified from the WSIs to measure the level of stain variation present within, and between, the eight laboratories. The laboratories used similar staining instruments and reagents, but their H&E stain durations varied. Intra-laboratory consistency was high (9-12% variation) but inter-laboratory variation was higher (maximum median difference of 31%) and the eight laboratories were statistically significantly different (p < 0.001). In addition, multiple colour-normalisation algorithms were applied to the image data using a range of target images. An automated nuclear count was then applied as an indicator of the impact of colour normalisation on the dataset and compared with pathologist manual 'ground truth' count. From three representative labs, the manual 'ground truth' nuclear counts ranged between 129 and 249, and the automated nuclear counts ranged between 152 and 621. Colour normalisation did improve inter-laboratory automated count variation (QCD) from 10% to 5%, but the results varied, particularly depending upon target image selection. The results from this work suggest that colour normalisation may improve consistency between slides but has the potential to be a confounder in its own right.

Hematoxylin and eosin (H&E) staining accounts for over 80% of slides stained worldwide. Although routinely used, there are high levels of variation between labs due to different staining methods. Staining is a pivotal part of slide preparation, but quality control is largely subjective, with overall clinical assurance provided by external quality assessment (EQA) services, underpinned by expert assessment. Digital pathology offers the potential to provide objective quantification of stain, through color analysis, to augment EQA assessment.This large-scale study evaluated H&E staining in 247 international labs participating in the UK NEQAS CPT EQA programme. Tissue sections were circulated to each lab to stain using their routine H&E staining protocol. The slides were reviewed by independent expert UK NEQAS CPT assessors, and quantitative digital analysis was conducted, comprising of H&E color deconvolution and color difference determination (ΔE).Most labs (69%) achieved an EQA score indicating good or excellent staining, with high inter-observer concordance to support this (92.5% within one mark of each other). H&E color difference, ΔE, showed 60% of labs were within 2 ΔE of the mean, which is considered as only perceptible through close observation. There was little correlation found between H&E intensity and assessor score, however, the H&E intensity ratio indicated a trend with assessor score suggesting there may be an optimal stain relationship that should be investigated further.The presented hybrid analysis combines expert analysis with objective data. This has the potential to inform upon optimal tissue staining and allows us to consider quantitative standards of H&E staining in pathology practice.

Whole slide imaging (WSI) of pathology glass slides with high-resolution scanners has enabled the large-scale application of artificial intelligence (AI) in pathology, to support the detection and diagnosis of disease, potentially increasing efficiency and accuracy in tissue diagnosis. Despite the promise of AI, it has limitations. 'Brittleness' or sensitivity to variation in inputs necessitates that large amounts of data are used for training. AI is often trained on data from different scanners but not usually by replicating the same slide across scanners. The utilisation of multiple WSI instruments to produce digital replicas of the same glass slides will make more comprehensive datasets and may improve the robustness and generalisability of AI algorithms as well as reduce the overall data requirements of AI training. To this end, the National Pathology Imagine Cooperative (NPIC) has built the AI FORGE (Facilitating Opportunities for Robust Generalisable data Emulation), a unique multi-scanner facility embedded in a clinical site in the NHS to (a) compare scanner performance and (b) replicate digital pathology image datasets across WSI systems. The NPIC AI FORGE currently comprises 15 scanners from 9 manufacturers. It can generate approximately 4000 WSI images per day (approximately 7Tb of image data). This paper describes the process followed to plan and build such a facility.

Staining tissue samples to visualise cellular detail and tissue structure is at the core of pathology diagnosis, but variations in staining can result in significantly different appearances of the tissue sample. While the human visual system is adept at compensating for stain variation, with the growth of digital imaging in pathology, the impact of this variation can be more profound. Despite the ubiquity of haematoxylin and eosin staining in clinical practice worldwide, objective quantification is not yet available. We propose a method for quantitative haematoxylin and eosin stain assessment to facilitate quality assurance of histopathology staining, enabling truly quantitative quality control and improved standardisation. The stain quantification method comprises conventional microscope slides with a stain-responsive biopolymer film affixed to one side, called stain assessment slides. The stain assessment slides were characterised with haematoxylin and eosin, and implemented in one clinical laboratory to quantify variation levels. Stain assessment slide stain uptake increased linearly with duration of haematoxylin and eosin staining (r = 0.99), and demonstrated linearly comparable staining to samples of human liver tissue (r values 0.98–0.99). Laboratory implementation of this technique quantified intra- and inter-instrument variation of staining instruments at one point in time and across a five-day period. The proposed method has been shown to reliably quantify stain uptake, providing an effective laboratory quality control method for stain variation. This is especially important for whole slide imaging and the future development of artificial intelligence in digital pathology.

Background:Currently, there is a paucity of guidelines relating to displays used for digital pathology making procurement decisions, and optimal display configuration, challenging.Experience suggests pathologists have personal preferences for brightness when using a conventional microscope which we hypothesized could be used as a predictor for display setup.Methods:We conducted an online survey across six NHS hospitals, totalling 108 practicing pathologists, to capture brightness adjustment habits on both microscopes and displays.A convenience subsample of respondents was then invited to take part in a practical task to determine microscope brightness and display luminance preferences in the normal working environment. A novel adaptation for a lightmeter was developed to directly measure the light output from the microscope eyepiece.Results:The survey (response rate 59% n=64) indicates 81% of respondents adjust the brightness on their microscope. In comparison, only 11% report adjusting their digital display. Display adjustments were more likely to be for visual comfort and ambient light compensation rather than for tissue factors, common for microscope adjustments. Part of this discrepancy relates to lack of knowledge of how to adjust displays and lack of guidance on whether this is safe; But, 66% felt that the ability to adjust the light on the display was important.Twenty consultants took part in the practical brightness assessment. Light preferences on the microscope showed no correlation with display preferences, except where a pathologist has a markedly brighter microscope light preference. All of the preferences in this cohort were for a display luminance of <500 cd/m2, with 90% preferring 350 cd/m2 or less. There was no correlation between these preferences and the ambient lighting in the room.Conclusions:We conclude that microscope preferences can only be used to predict display luminance requirements where the microscope is being used at very high brightness levels. A display capable of a brightness of 500 cd/m2 should be suitable for almost all pathologists with 300 cd/m2 suitable for the majority. Although display luminance is not frequently changed by users, the ability to do so was felt to be important by the majority of respondents.Further work needs to be undertaken to establish the relationship between diagnostic performance, luminance preferences, and ambient lighting levels.

Background: Pathology services experienced a surge in demand during the COVID-19 pandemic. Digitalisation of pathology workflows can help to increase throughput, yet many existing digitalisation solutions use non-standardised workflows captured in proprietary data formats and processed by black-box software, yielding data of varying quality. This study presents the views of a UK-led expert group on the barriers to adoption and the required input of measurement science to improve current practices in digital pathology. Methods: With an aim to support the UK’s efforts in digitalisation of pathology services, this study comprised: (1) a review of existing evidence, (2) an online survey of domain experts, and (3) a workshop with 42 representatives from healthcare, regulatory bodies, pharmaceutical industry, academia, equipment, and software manufacturers. The discussion topics included sample processing, data interoperability, image analysis, equipment calibration, and use of novel imaging modalities. Findings: The lack of data interoperability within the digital pathology workflows hinders data lookup and navigation, according to 80% of attendees. All participants stressed the importance of integrating imaging and non-imaging data for diagnosis, while 80% saw data integration as a priority challenge. 90% identified the benefits of artificial intelligence and machine learning, but identified the need for training and sound performance metrics.Methods for calibration and providing traceability were seen as essential to establish harmonised, reproducible sample processing, and image acquisition pipelines. Vendor-neutral data standards were seen as a “must-have” for providing meaningful data for downstream analysis. Users and vendors need good practice guidance on evaluation of uncertainty, fitness-for-purpose, and reproducibility of artificial intelligence/machine learning tools. All of the above needs to be accompanied by an upskilling of the pathology workforce. Conclusions: Digital pathology requires interoperable data formats, reproducible and comparable laboratory workflows, and trustworthy computer analysis software. Despite high interest in the use of novel imaging techniques and artificial intelligence tools, their adoption is slowed down by the lack of guidance and evaluation tools to assess the suitability of these techniques for specific clinical question. Measurement science expertise in uncertainty estimation, standardisation, reference materials, and calibration can help establishing reproducibility and comparability between laboratory procedures, yielding high quality data and providing higher confidence in diagnosis.

Background: The COVID-19 pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has placed a significant demand on healthcare providers (HCPs) to provide respiratory support for patients with moderate to severe symptoms. Continuous Positive Airway Pressure (CPAP) non-invasive ventilation can help patients with moderate symptoms to avoid the need for invasive ventilation in intensive care. However, existing CPAP systems can be complex (and thus expensive) or require high levels of oxygen, limiting their use in resource-stretched environments.Technical Development + Testing: The LeVe (“Light”) CPAP system was developed using principles of frugal innovation to produce a solution of low complexity and high resource efficiency. The LeVe system exploits the air flow dynamics of electric fan blowers which are inherently suited to delivery of positive pressure at appropriate flow rates for CPAP. Laboratory evaluation demonstrated that performance of the LeVe system was equivalent to other commercially available systems used to deliver CPAP, achieving a 10 cm H2O target pressure within 2.4% RMS error and 50–70% FiO2 dependent with 10 L/min oxygen from a commercial concentrator.Pilot Evaluation: The LeVe CPAP system was tested to evaluate safety and acceptability in a group of ten healthy volunteers at Mengo Hospital in Kampala, Uganda. The study demonstrated that the system can be used safely without inducing hypoxia or hypercapnia and that its use was well-tolerated by users, with no adverse events reported.Conclusions: To provide respiratory support for the high patient numbers associated with the COVID-19 pandemic, healthcare providers require resource efficient solutions. We have shown that this can be achieved through frugal engineering of a CPAP ventilation system, in a system which is safe for use and well-tolerated in healthy volunteers. This approach may also benefit other respiratory conditions which often go unaddressed in Low and Middle Income Countries (LMICs) for want of context-appropriate technology designed for the limited oxygen resources available.

There has been a marked rise in the number of avoidable deaths in health services around the world. At the same time there has been a growing increase in antibiotic resistant so-called "superbugs." We examine here the potential role of body temperature measurement in these adverse trends. Electronic based thermometers have replaced traditional mercury (and other liquid-in-glass type) thermometers for reasons of safety rather than superiority. Electronic thermometers are in general less robust from a measurement perspective than their predecessors. We illustrate the implications of unreliable temperature measurement on the diagnosis and management of disease, including COVID-19, through statistical calculations. Since a return to mercury thermometers is both undesirable and impractical, we call for better governance in the current practice of clinical thermometry to ensure the traceability and long-term accuracy of electronic thermometers and discuss how this could be achieved.

Pathology departments must rise to new staffing challenges caused by the coronavirus disease-19 pandemic and may need to work more flexibly for the foreseeable future. In light of this, many pathologists and departments are considering the merits of remote or home reporting of digital cases. While some individuals have experience of this, little work has been done to determine optimum conditions for home reporting, including technical and training considerations. In this publication produced in response to the pandemic, we provide information regarding risk assessment of home reporting of digital slides, summarize available information on specifications for home reporting computing equipment, and share access to a novel point-of-use quality assurance tool for assessing the suitability of home reporting screens for digital slide diagnosis. We hope this study provides a useful starting point and some practical guidance in a difficult time. This study forms the basis of the guidance issued by the Royal College of Pathologists, available at: https://www.rcpath.org/uploads/assets/626ead77-d7dd-42e1-949988e43dc84c97/RCPath-guidance-for-remote-digital-pathology.pdf.

Abstract Purpose: As pathology departments around the world contemplate digital microscopy for primary diagnosis, making an informed choice regarding display procurement is very challenging in the absence of defined minimum standards. In order to help inform the decision, we aimed to conduct an evaluation of displays with a range of technical specifications and sizes. Approach: We invited histopathologists within our institution to take part in a survey evaluation of eight short-listed displays. Pathologists reviewed a single haematoxylin and eosin whole slide image of a benign nevus on each display and gave a single score to indicate their preference in terms of image quality and size of the display. Results: Thirty-four pathologists took part in the display evaluation experiment. The preferred display was the largest and had the highest technical specifications (11.8-MP resolution, 2100  cd  /  m2 maximum luminance). The least preferred display had the lowest technical specifications (2.3-MP resolution, 300  cd  /  m2 maximum luminance). A trend was observed toward an increased preference for displays with increased luminance and resolution. Conclusions: This experiment demonstrates a preference for large medical-grade displays with the high luminance and high resolution. As cost becomes implicated in procurement, significantly less expensive medical-grade displays with slightly lower technical specifications may be the most cost-effective option.

Pathology services are facing pressures due to the COVID-19 pandemic. Digital pathology has the capability to meet some of these unprecedented challenges by allowing remote diagnoses to be made at home, during periods of social distancing or self-isolation. However, while digital pathology allows diagnoses to be made on standard computer screens, unregulated home environments may not be conducive for optimal viewing conditions. There is also a paucity of experimental evidence available to support the minimum display requirements for digital pathology. This study presents a Point-of-Use Quality Assurance (POUQA) tool for remote assessment of viewing conditions for reporting digital pathology slides. The tool is a psychophysical test combining previous work from successfully implemented quality assurance tools in both pathology and radiology to provide a minimally intrusive display screen validation task, before viewing digital slides. The test is specific to pathology assessment in that it requires visual discrimination between colors derived from hematoxylin and eosin staining, with a perceptual difference of ±1 delta E (dE). This tool evaluates the transfer of a 1 dE signal through the digital image display chain, including the observers’ contrast and color responses within the test color range. The web-based system has been rapidly developed and deployed as a response to the COVID-19 pandemic and may be used by anyone in the world to help optimize flexible working conditions at: http://www. virtualpathology.leeds.ac.uk/res earch/systems/pouqa/.

Here we report an investigation to adapt existing non-invasive ventilators (NIV) capable of delivering CPAP for use with oxygen to deliver enriched ventilation of 40%+ FiO2. Our intention is to maximise use of existing resources available to HCPs, as NIV and sleep apnoea (CPAP) machines are widely available, to deliver therapeutic benefit and potentially avoid the need for positive pressure ventilation during the COVID-19 pandemic.

Context.-Flexible working at diverse or remote sites is a major advantage when reporting using digital pathology, but currently there is no method to validate the clinical diagnostic setting within digital microscopy. Objective.-To develop a preliminary Point-of-Use Quality Assurance (POUQA) tool designed specifically to validate the diagnostic setting for digital microscopy. Design.-We based the POUQA tool on the red, green, and blue (RGB) values of hematoxylin-eosin. The tool used 144 hematoxylin-eosin-colored, 5x5-cm patches with a superimposed random letter with subtly lighter RGB values from the background color, with differing levels of difficulty. We performed an initial evaluation across 3 phases within 2 pathology departments: 1 in the United Kingdom and 1 in Sweden. Results.-In total, 53 experiments were conducted across all phases resulting in 7632 test images viewed in all. Results indicated that the display, the user's visual system, and the environment each independently impacted performance. Performance was improved with reduction in natural light and through use of medical-grade displays. Conclusions.-The use of a POUQA tool for digital microscopy is essential to afford flexible working while ensuring patient safety. The color-contrast test provides a standardized method of comparing diagnostic settings for digital microscopy. With further planned development, the color-contrast test may be used to create a "Verified Login" for diagnostic setting validation.

Digital microscopy produces high resolution digital images of pathology slides. Because no acceptable and effective control of color reproduction exists in this domain, there is significant variability in color reproduction of whole slide images. Guidance from international bodies and regulators highlights the need for color standardization. To address this issue, we systematically measured and analyzed the spectra of histopathological stains. This information was used to design a unique color calibration slide utilizing real stains and a tissue-like substrate, which can be stained to produce the same spectral response as tissue. By closely mimicking the colors in stained tissue, our target can provide more accurate color representation than film-based targets, whilst avoiding the known limitations of using actual tissue. The application of the color calibration slide in the clinical setting was assessed by conducting a pilot user-evaluation experiment with promising results. With the imminent integration of digital pathology into the routine work of the diagnostic pathologist, it is hoped that this color calibration slide will help provide a universal color standard for digital microscopy thereby ensuring better and safer healthcare delivery.

The crown of a human tooth has an outer layer of highly-mineralized tissue called enamel, beneath which is dentin, a less-mineralized tissue which forms the bulk of the tooth-crown and root. The composition and structure of enamel and dentin are different, resulting in different thermal properties. This gives an opportunity to characterize enamel and dentin from their thermal properties and to visually present the findings as a thermal map. The thermal properties of demineralized enamel and dentin may also be sufficiently different from sound tissue to be seen on a thermal map, underpinning future thermal assessment of caries. The primary aim of this novel study was to produce a thermal map of a sound, human tooth-slice to visually characterize enamel and dentin. The secondary aim was to map a human tooth-slice with demineralized enamel and dentin to consider future diagnostic potential of thermal maps for caries-detection. Two human slices of teeth, one sound and one demineralized from a natural carious lesion, were cooled on ice, then transferred to a hotplate at 30°C where the rewarming-sequence was captured by an infra-red thermal camera. Calculation of thermal diffusivity and thermal conductivity was undertaken, and two methods of data-processing used customized software to produce thermal maps from the thermal characteristic-time-to-relaxation and heat-exchange. The two types of thermal maps characterized enamel and dentin. In addition, sound and demineralized enamel and dentin were distinguishable within both maps. This supports thermal assessment of caries and requires further investigation on a whole tooth.

Introduction/ Background Guidance from the Food and Drug Administration has recommended color standardization for whole slide imaging, as with all other digital systems. However there is known, unresolved and substantial variation in color between digital slide scanners. To address this issue, we created a novel color calibration test object which uniquely utilizes histochemical stains and a tissue-like substrate, affording accurate internal calibration of WSIs. Aims We aimed to evaluate the clinical application of the novel test object. Objectives included: 1. Whether calibration made WSIs appear closer in color to the glass slide counterpart; 2. Whether pathologists prefer calibrated WSIs; and, 3. Whether calibration affected diagnostic confidence. Methods Six pathology cases that present known difficulties when viewed using a digital microscope were selected and WSIs were generated. These WSIs were calibrated using a color ICC profile created using spectral measurements from the test object. Twelve pathologists, blinded to intervention, compared calibrated and uncalibrated versions of each WSI on a medical-grade monitor. The display was color calibrated using a colorimeter which accounted for ambient lighting. Three, subjective responses were recorded on 7-point Likert scales. Results Calibrated WSIs were closer in appearance to the microscope in 40 of 72 trials, (56%) and were preferred in 46 of 72 trials (64%). Calibration improved diagnostic confidence (median 6.00 vs. 5.00, p=0.001). Diagnostic confidence with the calibrated slides was correlated with preference for color calibration (r=0.499; p<0.001).

Performing diagnoses using virtual slides can take pathologists significantly longer than with glass slides, presenting a significant barrier to the use of virtual slides in routine practice. Given the benefits in pathology workflow efficiency and safety that virtual slides promise, it is important to understand reasons for this difference and identify opportunities for improvement. The effect of display resolution on time to diagnosis with virtual slides has not previously been explored. The aim of this study was to assess the effect of display resolution on time to diagnosis with virtual slides. Nine pathologists participated in a counterbalanced crossover study, viewing axillary lymph node slides on a microscope, a 23-in 2.3-megapixel single-screen display and a three-screen 11-megapixel display consisting of three 27-in displays. Time to diagnosis and time to first target were faster on the microscope than on the single and three-screen displays. There was no significant difference between the microscope and the three-screen display in time to first target, while the time taken on the single-screen display was significantly higher than that on the microscope. The results suggest that a digital pathology workstation with an increased number of pixels may make it easier to identify where cancer is located in the initial slide overview, enabling quick location of diagnostically relevant regions of interest. However, when a comprehensive, detailed search of a slide has to be made, increased resolution may not offer any additional benefit.